Heterologous abs from llama and chicken egg yolk to prevent rotavirus diarrhea

描述(申请人提供)：我们将探索口服不同的被动异源抗体(Abs)、蛋黄IgY和来自骆驼重链Abs的重组VHH片段，作为轮状病毒(RV)腹泻预防和治疗疫苗接种的替代或补充方法。被动治疗将在两种轮状病毒病的新生动物模型上进行评估，GnotoBiotic(Gn)猪和牛犊。这项研究将同时在美国的赛义夫博士实验室、俄亥俄州立大学(Gn PIG)和阿根廷的INTA与Parreqo博士(新生小牛)合作进行，作为NIAID的延伸，NIH授予编号。R01 AI033561-11(6/25/03-12/31/08)赛义夫博士与Parreqo博士和Fernandez博士(INTA)之间持续和长期的合作促进了最先进技术的转让，用于诊断(分析、试剂)和治疗(疫苗)牛的轮状病毒感染，牛是阿根廷的主要农产品。通过这项建议，我们通过探索新的免疫学概念(被动异源抗体对RV主动抗体反应的影响)和新的治疗方法(被动IgY，VHH)，在两种易患RV腹泻的新生动物：具有正常肠道微生物菌群的常规小牛和缺乏微生物菌群的Gn猪中，继续开展与改善反刍动物和人类健康相关的合作。国外合作者将生产抗体(骆驼VHH、蛋黄抗体、牛免疫球蛋白)，并进行小牛试验。INTA的学生将在赛义夫博士的实验室接受培训，学习新的免疫学程序(流式细胞术等)，使用专门的动物疾病模型(Gn猪)，以及提高与粘膜免疫相关的分析技能。因此，INTA团队将接受与研究Gn猪粘膜免疫反应相关的试剂和培训，这些试剂和培训将应用于小牛实验。我们的合作还将进一步提高INTA作为拟议领域基础研究和翻译研究卓越中心的声誉。从大羊驼中开发重组抗体，将使这种本地动物能够可持续地应用于VHH抗体的开发，用于治疗这种疾病和其他疾病。这将有利于骆驼畜牧业，以及阿根廷或南美洲其他较贫穷的发展中国家(秘鲁、玻利维亚等)从事骆驼生产的区域微观经济。这些异源被动抗体治疗将对早产儿或在严重的轮状病毒暴发环境(医院、日托中心、发展中国家的农村地区)产生深远的影响。它们代表了在不可能、太昂贵、太迟使用轮状病毒疫苗的情况下减少轮状病毒腹泻严重程度和死亡的补充治疗方法；或者用于治疗婴儿和反刍动物疫苗失败的病例。公共卫生相关性：轮状病毒(RV)是全球幼小动物和人类婴儿腹泻的主要原因。目前，轮状病毒在动物中的预防是基于灭活的轮状病毒母体疫苗以增强被动免疫，而减毒活疫苗最近已被批准用于人类婴儿。疫苗的效力和安全性仍不清楚，特别是在发展中国家。我们将探索口服不同的被动异源抗体、骆驼重链抗体片段(VHH)和蛋黄IgY作为替代/互补方法来控制轮状病毒感染和疾病识别(Gn)猪和新生牛犊两种动物模型中的轮状病毒腹泻。如果我们的异源被动抗体(Ab)策略有效，这些治疗方法将产生最深远的影响，作为早产儿或严重RV爆发环境(医院、日托中心、发展中国家农村地区)的补充剂，通过提供替代的快速治疗方法，在不可能、太昂贵、太晚使用RV疫苗的情况下降低RV腹泻的严重性和死亡率；或治疗疫苗失败的病例。

DESCRIPTION (provided by applicant): We will explore oral administration of different passive heterologous antibodies (Abs), chicken egg yolk IgY and recombinant VHH fragments derived from llama heavy chain Abs as an alternative or complementary approach to vaccination for rotavirus (RV) diarrhea prevention and treatment. The passive treatments will be evaluated in two neonatal animal models of RV disease, gnotobiotic (Gn) pigs and calves. The research will be done simultaneously, in the US at Dr. Saif's Lab, The Ohio State University (Gn pig) and in Argentina, at INTA in collaboration with Dr. Parreqo (neonatal calves), as an extension of NIAID, NIH Grant No. R01 AI033561-11 (6/25/03-12/31/08). The ongoing and long-term collaboration between Dr. Saif and Drs. Parreqo and Fernandez (INTA) has facilitated transfer of state-of-the-art technology for diagnosis (assays, reagents) and treatments (vaccines) for RV infections of cattle, the major agricultural commodity in Argentina. Through this proposal we continue this collaboration related to improving both ruminant and human health by exploring new immunologic concepts (impact of passive heterologous Ab on active Ab responses to RV) and new treatments (passive IgY,VHH) in two neonatal animals susceptible to RV diarrhea: conventional calves with a normal gut microbial flora and Gn pigs lacking the microbial flora. The foreign collaborator will produce the Abs (llama VHH, chicken egg IgY, bovine IgG) and conduct the calf experiments. Students from INTA will be trained in Dr. Saif's lab to learn newer immunologic procedures (flow cytometry, etc) and work with specialized animal disease models (Gn pigs) as well as to enhance analytical skills related to mucosal immunity. Thus the INTA team will receive reagents and training related to the study of mucosal immune responses in Gn pigs that will be applied to the calf experiments. Our collaboration will also further INTA's reputation as a Center of Excellence for basic and translational studies in the proposed areas. Development of recombinant Abs from llamas, will enable a sustainable application of this native animal for the development of VHH Ab applied to treat this and other diseases. This will benefit llama livestock production, as well as regional micro-economies for Argentina or other poorer developing countries of South America (Peru, Bolivia, etc) engaged in Camelid production. These heterologous passive Ab treatments would have profound impacts as supplements for premature infants or in severe RV outbreak settings (hospitals, day care centers, rural areas of developing countries). They represent complementary treatments to reduce RV diarrhea severity and deaths under conditions where RV vaccine use is not possible, too expensive, too late; or to treat cases of vaccine failure for both infants and ruminants. PUBLIC HEALTH RELEVANCE: Rotavirus (RV) is the leading cause of diarrhea in young animals and human infants, worldwide. Currently, RV prevention in animals is based on inactivated RV maternal vaccines to enhance passive immunity, whereas live-attenuated RV vaccines have been recently approved for human infants. Vaccine efficacy and safety remain unclear especially in developing countries. We will explore oral administration of different passive heterologous antibodies, llama heavy chain antibody fragments (VHH) and chicken egg yolk IgY as alternative/complementary approaches to control RV diarrhea for both animals and humans, in two animal models of rotavirus infection and disease gnotobiotic (Gn) pigs and neonatal calves. If our heterologous passive antibody (Ab) strategies are effective, these treatments would have the most profound impacts as supplements for premature infants or in severe RV outbreak settings (hospitals, day care centers, rural areas of developing countries), by providing an alternative rapid treatment to reduce RV diarrhea severity and deaths under conditions where RV vaccine use is not possible, too costly, too late; or to treat cases of vaccine failure.